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Recreating Human Heart Development in the Lab

Recreating Human Heart Development in the Lab | iBB | Scoop.it

The embryonic development of the human heart under healthy and pathological conditions is still very poorly understood. In a recently published Nature Communications paper, a iBB team led by Margarida Diogo, in collaboration with Perpetua Pinto do Ó at i3S, engineered a unique organ-like model from stem cells that recreates several stages of embryonic development of the human heart. This humanized model opens now the path to study, in a tissue engineering lab, the development of congenital heart diseases, such as coronary vascular diseases and myocardium non-compaction cardiomyopathies, and can be further applied in drug screening and toxicology assays during embryonic heart development. This last topic is particularly relevant since pregnant women normally do not take part in clinical trials, making this human cell-based in vitro platform the only viable alternative to assess developmental cardiotoxicity. The work was a part of the PhD thesis in Bioengineering – MIT Portugal program of Mariana Branco, the first author of the paper, and was funded by the FCT project “Mini-Hearts - Organoid Engineering for Production of 3D Cardiovascular Microtissues from Human Induced Pluripotent Stem Cells for Cardiotoxicity”.  

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Modeling Rett Syndrome With Human Patient-Specific Forebrain Organoids

Modeling Rett Syndrome With Human Patient-Specific Forebrain Organoids | iBB | Scoop.it

Engineering brain organoids from human induced pluripotent stem cells (hiPSCs) is a powerful tool for modeling brain development and neurological disorders. Rett syndrome (RTT), a rare neurodevelopmental disorder, can greatly benefit from this technology, since it affects multiple neuronal subtypes in forebrain sub-regions. SCERG-iBB researchers have recently established dorsal and ventral forebrain organoids from control and RTT patient-specific hiPSCs recapitulating the 3D organization and functional network complexity of this brain region. The data obtained revealed a premature development of the deep-cortical layer, associated to the formation of TBR1 and CTIP2 neurons, and a lower expression of neural progenitor/proliferative cells in RTT dorsal organoids. Moreover, calcium imaging and electrophysiology analysis demonstrated functional defects of RTT neurons. Additionally, assembly of RTT dorsal and ventral organoids revealed impairments of interneuron’s migration. Overall, these models provide a better understanding of RTT during early stages of neural development, demonstrating a great potential for personalized diagnosis and drug screening. The paper was published in Frontiers in Cell Development Biology.

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Transcriptomic Analysis of 3D Cardiac Differentiation of Human Induced Pluripotent Stem Cells

Transcriptomic Analysis of 3D Cardiac Differentiation of Human Induced Pluripotent Stem Cells | iBB | Scoop.it

Human induced pluripotent stem cells (hiPSCs) represent an almost limitless source of cells for biomedical applications including cardiomyocytes (CMs), the most predominant cell type in the human heart. iBB researchers have established an efficient and robust 3D platform for CM production from hiPSCs and studied the impact of 3D culture on CM differentiation and maturation compared with a 2D monolayer culture. It was found that CMs mature earlier and show an improved communication system in this 3D environment which was suggested to be responsible for a higher structural and functional maturation. This novel 3D culture platform and the CMs obtained can be used for disease modelling, drug screening and cardiotoxicity tests. The results were published in Scientific Reports.

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Neural Induction of Human Induced Pluripotent Stem Cells for Neurodevelopmental Toxicity Studies

Neural Induction of Human Induced Pluripotent Stem Cells for Neurodevelopmental Toxicity Studies | iBB | Scoop.it

The ability to differentiate neural progenitors (NP) from human induced pluripotent stem cells (hiPSCs) provides an opportunity to develop new applications for cellular therapy, disease modelling and drug screening. SCERG-iBB researchers developed a platform that can be applied towards the study of the effect of neurotoxic molecules that impair normal embryonic development, such as the antiepileptic drug valproic acid (VPA). It was verified that exposure to VPA led to a prevalence of NP structures over neuronal differentiation, confirmed by analysis of the expression of neural cell adhesion molecule, and neural rosette number and morphology. This methodology can potentially complement current toxicity tests for the detection of teratogenic compounds that can interfere with normal embryonic development. The work was published in Toxicology Letters.

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3D Differentiation of Human Oligodendrocyte Precursors from Pluripotent Stem Cells

3D Differentiation of Human Oligodendrocyte Precursors from Pluripotent Stem Cells | iBB | Scoop.it
Oligodendrocyte precursor cells (OPCs) offer considerable potential for the treatment of demyelinating diseases and injuries of the CNS. However, generating large quantities of high-quality OPCs remains a challenge that impedes their therapeutic application. In collaboration with researchers at the Stem Cell Engineering Research Group (SCERG), researchers at the Department of Bioengineering, University of California, Berkeley, have recently developed a novel defined and scalable culture system for differentiation of OPCs from Human Pluripotent Stem Cells, under 3D conditions, using a fully-defined thermoresponsive biomaterial. This system enabled the generation of large quantities of high-quality OPCs that were able to engraft, migrate and mature into myelinating oligodendrocytes upon transplantation into the brains of NOD/SCID mice.
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Modeling Rett Syndrome with Human Pluripotent Stem Cells: Mechanistic Outcomes and Future Clinical Perspectives

Modeling Rett Syndrome with Human Pluripotent Stem Cells: Mechanistic Outcomes and Future Clinical Perspectives | iBB | Scoop.it

Rett syndrome (RTT) is a rare neurodevelopmental disorder caused by mutations in the gene encoding for the MeCP2 protein. Among different roles, MeCP2 has a high phenotypic impact during the different stages of brain development. Thus, it is essential to investigate the function of MeCP2 and its regulated targets. In a review paper published in the International Journal of Molecular Sciences, a team of researchers at SCERG-iBB provides a brief summary of the main neurological features of RTT and of the impact of MeCP2 mutations in the neuropathophysiology of the disease. A thorough revision of recent advances and future prospects of RTT modeling using human neural cells derived from pluripotent stem cells and its contribution for the current and future clinical trials for RTT is also provided.

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Maturation of Human Pluripotent Stem Cell-Derived Cerebellar Neurons in the Absence of Co-Culture

Maturation of Human Pluripotent Stem Cell-Derived Cerebellar Neurons in the Absence of Co-Culture | iBB | Scoop.it

In a new paper published in Frontiers in Bioengineering and Biotechnology, SCERG-iBB researchers in collaboration with colleagues from the Institute of Molecular Medicine (iMM) describe a novel differentiation strategy that uses defined medium to generate Purkinje cells, granule cells, interneurons, and deep cerebellar nuclei projection neurons, that self-formed and matured into electrically active cells. This research is expected to result in better models for the study of cerebellar dysfunctions and represent an important advancement towards the development of autologous replacement strategies for treating cerebellar degenerative diseases.

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Expansion of Human Induced Pluripotent Stem Cells in Vertical-Wheel Bioreactors

Expansion of Human Induced Pluripotent Stem Cells in Vertical-Wheel Bioreactors | iBB | Scoop.it

The successful use of Human induced Pluripotent Stem Cells (hiPSC) for disease modelling, drug discovery and, ultimately, for regenerative therapies depends on the development of robust bioprocesses capable of generating large numbers of hiPSC and derivatives. SCERG-iBB researchers developed a bioprocess for the scalable generation of hiPSC in a microcarrier-based system using, for the first time, single-use Vertical-Wheel bioreactors. hiPSC culture was performed in working volumes up to 300 mL, maintaining the pluripotency and genomic integrity of the cells, providing an important tool for the successful manufacturing of hiPSC-based products.  The work was published in the Journal of Chemical Technology and Biotechnology.

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Structural Maturation of Human Cardiomyocytes Derived from Pluripotent Stem Cells

Structural Maturation of Human Cardiomyocytes Derived from Pluripotent Stem Cells | iBB | Scoop.it

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) have an enormous potential for in vitro modeling of cardiac diseases and for testing the effect and toxicity of new drugs. However, when compared to adult CMs, hiPSC-CMs are very immature, exhibiting low structural development and functionality. SCERG-iBB researchers developed a novel methodological framework to quantify structural aspects of hiPSC-CMs during long-term culture. hiPSC-CMs showed significant progression in several structural characteristics namely cardiomyocyte fiber density and length. Importantly, this methodology contributes to set new metrics to develop applications for drug screening and disease modeling for hiPSC-CMs. The research has been published on Biochemical and Biophysical Research Communications.

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